An endoribonuclease-based feedforward controller for decoupling resource-limited genetic modules in mammalian cells.
An endoribonuclease-based feedforward controller for decoupling resource-limited genetic modules in mammalian cells.
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DOI：
10.1038/s41467-020-19126-9
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发表时间：
2020-11-10
影响因子：
16.6
通讯作者：
Del Vecchio D
中科院分区：
文献类型：
--
作者：
Jones RD;Qian Y;Siciliano V;DiAndreth B;Huh J;Weiss R;Del Vecchio D
合成生物学有潜力为医疗保健和生物技术的应用带来先进的遗传装置。然而，由于缺乏模块化，准确预测工程基因设备的行为仍然很困难，其中设备的输出不仅取决于其预期的输入，还取决于其环境。缺乏模块化的一个原因是转录和翻译资源的负载，这可以诱导其他独立调控的基因之间的偶联。在这里，我们量化了资源负荷在工程哺乳动物遗传系统中的影响，并开发了一种基于核糖核酸内切酶的前馈控制器，该控制器可以使感兴趣的基因的表达水平适应哺乳动物细胞中显著的资源负荷。核糖核酸内切酶的高产量和催化速率促进了对资源负荷的近乎完美的适应。我们的设计是可移植的跨细胞系，使控制器功能可预测的调整。最终，我们的控制器是一种通用设备，用于可预测的、健壮的和与上下文无关的基因表达控制。由于缺乏模块化，准确预测遗传电路的行为仍然很困难。在这里，作者量化了哺乳动物系统中资源负荷的影响，并开发了一种基于核糖核酸内切酶的前馈控制器，使基因表达适应资源负荷的影响。
Synthetic biology has the potential to bring forth advanced genetic devices for applications in healthcare and biotechnology. However, accurately predicting the behavior of engineered genetic devices remains difficult due to lack of modularity, wherein a device’s output does not depend only on its intended inputs but also on its context. One contributor to lack of modularity is loading of transcriptional and translational resources, which can induce coupling among otherwise independently-regulated genes. Here, we quantify the effects of resource loading in engineered mammalian genetic systems and develop an endoribonuclease-based feedforward controller that can adapt the expression level of a gene of interest to significant resource loading in mammalian cells. Near-perfect adaptation to resource loads is facilitated by high production and catalytic rates of the endoribonuclease. Our design is portable across cell lines and enables predictable tuning of controller function. Ultimately, our controller is a general-purpose device for predictable, robust, and context-independent control of gene expression. Accurately predicting the behaviour of a genetic circuit remains difficult due to the lack of modularity. Here the authors quantify the effects of resource loading in mammalian systems and develop an endoribonuclease-based feedfoward controller to adapt gene expression to the effects of resource loading.
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影响因子：
48
作者：
Chavez A;Tuttle M;Pruitt BW;Ewen-Campen B;Chari R;Ter-Ovanesyan D;Haque SJ;Cecchi RJ;Kowal EJK;Buchthal J;Housden BE;Perrimon N;Collins JJ;Church G
通讯作者：
Church G
影响因子：
14.9
作者：
Baron, U;Gossen, M;Bujard, H
通讯作者：
Bujard, H
影响因子：
16.6
作者：
Frei, Timothy;Cella, Federica;Siciliano, Velia
通讯作者：
Siciliano, Velia
影响因子：
16.6
作者：
Donahue, Patrick S.;Draut, Joseph W.;Leonard, Joshua N.
通讯作者：
Leonard, Joshua N.
影响因子：
2.2
作者：
Fuhrer, D.;Han, S.;Ludgate, M.
通讯作者：
Ludgate, M.